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This work is focused on the synthesis of leucyl-glycine-functionalized γ-Fe₂O₃ magnetic nanoparticles coated by polydioxanone (γ-Fe₂O₃-CA-Leu-Gly-PDX) as a polyethersulphone (PES) membrane for biotechnological applications. The physicochemical characteristics were investigated by FT-IR, SEM, XRD, a vibrating sample magnetometer (VSM), and ICP-OES. The present investigation also centered on the several biological activities of γ-Fe₂O₃-CA-Leu-Gly-PDX. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and metal chelating activity was studied for evaluation of its antioxidant activity potential. It exhibited 100% DPPH radical scavenging and 93.33% metal chelating activity. With applicability to antimicrobial photodynamic therapy, DNA cleavage and antimicrobial activity, the cell viability of γ-Fe₂O₃-CA-Leu-Gly-PDX was investigated in detail. The γ-Fe₂O₃-CA-Leu-Gly-PDX demonstrated the significant biofilm inhibition activity as being 81.54% and 86.34% for <i>P. aeruginosa</i> and <i>S. aureus</i>, respectively. Moreover, a novel polyethersulphone nanocomposite membrane incorporated with γ-Fe₂O₃-CA-Leu-Gly-PDX was prepared. The performance of the γ-Fe₂O₃-CA-Leu-Gly-PDX-blended polyethersulphone (PES) membrane was investigated by measuring the antifouling and <i>E. coli</i> rejection. The nanocomposite membranes demonstrated remarkable antifouling properties in contrast with the pristine PES when BSA (bovine serum albumin) and <i>E. coli</i> were filtrated. A complete rejection was obtained by the composite membrane. After an application of the membrane study, the modified polyethersulphone (PES) membrane blended with γ-Fe₂O₃-CA-Leu-Gly-PDX removed 100% of the <i>E. coli.</i>